Cloning, Expression, and Characterization of a New Phytase from the Phytopathogenic Bacterium Pectobacterium wasabiae DSMZ 18074

文献类型： 外文期刊

第一作者： Shao, Na

作者： Shao, Na;Huang, Huoqing;Meng, Kun;Luo, Huiying;Wang, Yaru;Yang, Peilong;Yao, Bin

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期刊名称：JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：2.351； 五年影响因子：2.65 ）

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收录情况： SCI

摘要： The soft rot bacterium Pectobacterium wasabiae is an economically important pathogen of many crops. A new phytase gene, appA, was cloned from P. wasabiae by degenerate PCR and TAIL-PCR. The open reading frame of appA consisted of 1,302 bp encoding 433 amino acid residues, including 27 residues of a putative signal peptide. The mature protein had a molecular mass of 45 kDa and a theoretical pI of 5.5. The amino acid sequence contained the conserved active site residues RHGXRXP and HDTN of typical histidine acid phosphatases, and showed the highest identity of 48.5% to PhyM from Pseudomonas syringae. The gene fragment encoding the mature phytase was expressed in Escherichia coli BL21 (DE3), and the purified recombinant phytase had a specific activity of 1,072??47 U/mg for phytate substrate. The optimum pH and temperature for the purified phytase were pH 5.0 and 50oC, respectively. The Km value was 0.17 mM, with a Vmax of 1,714 mmol/min/mg. This is the first report of the identification and isolation of phytase from Pectobacterium.

分类号： Q93

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